Abstract | AIM: METHODS: Human pancreatic cancer cells Panc-1, Mia-paca2, Bxpc-3 and SW1990, infected with lentivirus, were analyzed by real-time polymerase chain reaction (PCR). Western blotting was used to examine XIAP protein levels, survivin and p-Akt to confirm the result of real-time PCR and determine the possible mechanism. The 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to measure IC₅₀ to determine chemosensitivity to the chemotherapeutic drugs 5-fluorouracil (5-FU) and gemcitabine. A colony assay, MTT assay and a tumorigenicity experiment were used to study cell proliferation in vitro and in vivo. Caspase-3/7 activity, 4',6-diamidino-2-phenylindole-staining and flow cytometric measurements were used to study apoptosis in SW1990 cells. RESULTS: XIAP proteins were found to be differentially expressed among pancreatic cancer cell lines Panc-1, Mia-paca2, Bxpc-3 and SW1990. Data of real-time PCR and Western blotting showed that XIAP was reduced persistently and markedly by lentivirus-mediated shRNA. Downregulation of XIAP by transfection with XIAP shRNA resulted in decreased p-Akt expression. XIAP shRNA also inhibited the growth of pancreatic cancer cells in vitro and in vivo, enhanced drug-induced apoptosis and increased chemosensitivity to 5-FU and gemcitabine. Results also suggest that inhibition of XIAP and subsequent p-Akt depletion may have an anti- tumor effect through attenuating the ability of cancer cells to survive. CONCLUSION: Lentivirus-mediated gene therapy is an attractive strategy in the treatment of pancreatic cancer and justifies the use of lentivirus in pancreatic cancer gene therapy studies.
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Authors | Chun Jiang, Xiao-Ping Yi, Hong Shen, Yi-Xiong Li |
Journal | World journal of gastroenterology
(World J Gastroenterol)
Vol. 18
Issue 23
Pg. 2956-65
(Jun 21 2012)
ISSN: 2219-2840 [Electronic] United States |
PMID | 22736919
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Antimetabolites, Antineoplastic
- BIRC5 protein, human
- Inhibitor of Apoptosis Proteins
- RNA, Small Interfering
- Survivin
- X-Linked Inhibitor of Apoptosis Protein
- XIAP protein, human
- Deoxycytidine
- Proto-Oncogene Proteins c-akt
- Caspase 3
- Caspase 7
- Fluorouracil
- Gemcitabine
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Topics |
- Antimetabolites, Antineoplastic
(pharmacology)
- Apoptosis
(drug effects)
- Caspase 3
(metabolism)
- Caspase 7
(metabolism)
- Cell Line, Tumor
- Cell Proliferation
- Deoxycytidine
(analogs & derivatives, pharmacology)
- Down-Regulation
- Drug Resistance, Neoplasm
(genetics)
- Fluorouracil
(pharmacology)
- Genetic Therapy
- Humans
- Inhibitor of Apoptosis Proteins
(drug effects, metabolism)
- Pancreatic Neoplasms
(genetics, metabolism, therapy)
- Proto-Oncogene Proteins c-akt
(drug effects, metabolism)
- RNA, Small Interfering
(therapeutic use)
- Survivin
- X-Linked Inhibitor of Apoptosis Protein
(genetics, metabolism)
- Gemcitabine
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